DICP OpenIR
Subject Area物理化学
SECRETORY EXPRESSION OF THE EXO-INULINASE FROM K. MARXIANUS CBS 6556 IN PICHIA PASTORIS
Zhang SF(张素芳); Yang F(杨帆); Wang Q(王倩); Zhao ZB(赵宗保)
Conference Name13th International Biotechnology Symposium and Exhibition
Conference Date2008-10-12
2008-10-12
Conference Place中国
PagesS309/2
ISSN0168-1656
Department十八室
Funding Organization中国科学院,中国工程院,中国教育部,中国科技部
AbstractInulin is linear oligosaccharides [1]. Although inulin can be hydrolyzed to monosaccharides non-enzymatically by acid treatment, inhibitory by-products produced that complicate downstream biological process. Therefore, it is pivotal to establish a cost-effective inulinase (EC 3.2.1.7) production. In this study, we isolated the DNA sequence encoding the mature peptide sequence of the exo-inulinase gene from Kluyveromyces marxianus CBS 6556 and expressed in Pichia pastoris X-33. The purified recombinant enzyme (reINU) gave a specific activity of 13100 U•mg−1, which was about 100-fold higher that reported for the wild type protein isolated from K. marxianus CBS6556 [2, 3]. Ferguson plot analysis showed that secretion expressed reINU was a 169 kDa dimer. Detailed analytical assays showed that the optimal temperature and pH for reINU were 60 oC and pH 4.0, respectively. It was found that hydrolysis activity was about 20% higher in the presence of Mn2+. reINU was stable for over 3 days at room temperature. When employed for hydrolysis of the juice of the fresh tuber (water/tuber, 1:1) of Helianthus tuberosus at 50 oC for 1 h, total reduced sugar reached 65 g•l−1, and 85% of the total sugar of the raw material was released based on ion chromatography analysis. In conclusion, we have developed an overproduction system for inulinase that can now be used as biocatalysts for fructose syrups industry and biomass conversion research.
Language中文
WOS IDWOS:000208979401216
Citation statistics
Document Type会议论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/113172
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhao ZB(赵宗保)
Recommended Citation
GB/T 7714
Zhang SF,Yang F,Wang Q,et al. SECRETORY EXPRESSION OF THE EXO-INULINASE FROM K. MARXIANUS CBS 6556 IN PICHIA PASTORIS[C],2008:S309/2.
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